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Mitochondrial Apoptosis Detection Fluorometric Kit, MitoCapture

based on 38 citations in multiple journalsMitochondrial Apoptosis Detection Fluorometric Kit, MitoCapture385 5
Distinguishes between apoptotic & healthy cells
Catalog #: K250
SKU-Size Size Price Qty
K250-25 25 assays
$165.00
K250-100 100 assays
$405.00
More Sizes Get Quote

Product Details

Detection Method Fluorescence microscopy and Flow cytometry: FITC (Ex/Em = 488/530 ± 30 nm); and (optional) PI (Em = 488/590 ± 42 nm)
Species Reactivity Mammalian
Applications Disruption of mitochondrial transmembrane potential is one of the earliest intracellular events that occur upon induction of apoptosis. The MitoCapture Mitochondrial Apoptosis Detection Kit provides a simple and sensitive in vitro assay for detecting the mitochondrial changes in apoptosis. It is highly sensive and detects apoptosis in living cells.
Features & Benefits • Simple one-step procedure; takes only 30 minutes
• Fast and convenient
Kit Components • MitoCapture Reagent
• Incubation Buffer
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Disruption of the mitochondrial transmembrane potential is one of the earliest intracellular events that occur following induction of apoptosis. The MitoCapture™ Apoptosis Detection Kit provides a simple, fluorescent-based method for distinguishing between healthy and apoptotic cells by detecting the changes in the mitochondrial transmembrane potential. The kit utilizes MitoCapture™, a cationic dye that fluoresces differently in healthy vs apoptotic cells. In healthy cells, MitoCapture accumulates and aggregates in the mitocondria, giving off a bright red fluorescence. In apoptotic cells, MitoCapture cannot aggregate in the mitochondria due to the altered mitochondrial transmembrane potential, and thus it remains in the cytoplasm in its monomer form, fluorescing green. The fluorescent signals can be easily detected by fluorescence microscopy using a band-pass filter (detects FITC and rhodamine) or analyzed by flow cytometry using FITC channel for green monomers (Ex/Em = 488/530+ 30 nm) and (optional) PI channel for red ggregates (Em = 488/590+ 42 nm).


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How long wil the fluorescence signal last?
The fluorescence signal might not be equally intense for 48 hours but the samples must be kept protected from light and the intensity should be measurable within the first 24 hours or so. The fluorescence quenches over time.
Would it work for tissue samples fixed in paraffin?
K250 works with live cells. If you wish to use paraffin-fixed tissue and look for apoptosis signals, I would suggest using the TUNEL assay kits instead: http://www.biovision.com/search/results.html?keywords=tunel&x=0&y=0
Can I use this kit to evaluate drug-induced mitochondrial dysfunction?
The Kit detects the changes in the mitochondrial trans-membrane potential and helps differentiate between healthy and apoptotic cells. If by “mitochondrial dysfunction” you mean change in mitochondrial membrane potential resulting in a leaky membrane, then yes you can measure that using this kit.
Can you use frozen tissues to isolate microsomes?
We recommend using fresh tissues. However, sections frozen immediately after isolation can work too. The frozen tissues should not have undergone multiple rounds of freeze and thaw.
Can I obtain not only microsome fraction but also cytosol fraction with your Microsome Isolation Kit (Cat.# K249-50)?
You can obtain cytosolic fractions from the S9 fraction. The S9 fraction was further centrifuged at >20,000 × g (preferably at 100, 000g) for 30 min to separate the microsomes and cytosolic fraction (Yoshihara and Ohta, 1998). Collect the supernatant at this step, that should be your cytosolic fraction.
Which method for the quantification of total protein amount in microsomes would you recommend?
The protein in the microsomal isolate can be quantified with a BCA assay kit.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact yopur regional BioVision distributor.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Maria Plataki, Mitochondrial Dysfunction in Aged Macrophages and Lung during Primary Streptococcus pneumoniae Infection is Improved with Pirfenidone. Sci Rep, Jan 2019;  30700745.
Espinoza, J. Luis et al. (2017) The simultaneous inhibition of the mTOR and MAPK pathways with Gnetin-C induces apoptosis in acute myeloid leukemia, Cancer Lett. 2017 Aug 1;400:127-136.
Guan et al., Protective role of cyclosporine A and minocycline on mitochondrial disequilibrium-related podocyte injury and proteinuria occurrence induced by Adriamycin. Nephrol. Dial. Transplant., Jun 2015; 30: 957 - 969.
Wu et al., Selenoprotein H Suppresses Cellular Senescence through Genome Maintenance and Redox Regulation. J. Biol. Chem., Dec 2014; 289: 34378 - 34388.
DeNicola et al., Stimulation of glucagon-like peptide-1 receptor through exendin-4 preserves myocardial performance and prevents cardiac remodeling in infarcted myocardium. Am J Physiol Endocrinol Metab, Oct 2014; 307: E630 - E643.
For more citations of this product click here